by Lau Chun Hong. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2004. / Includes bibliographical references (leaves 179-195). / Abstracts in English and Chinese. / Abstract --- p.i / Abstract in Chinese 摘要 --- p.iv / Acknowledgements --- p.vi / Table of Contents --- p.vii / List of Abbreviations --- p.xiii / List of Figures --- p.xvi / List of Tables --- p.xviii / Publications --- p.xix / Chapter Chapter1 --- Introduction --- p.1 / Chapter 1.1 --- Epidemiology of Diabetes Mellitus --- p.1 / Chapter 1.2 --- Definition of Diabetes Mellitus --- p.2 / Chapter 1.3 --- Glucose Homeostasis and Diabetes Mellitus --- p.2 / Chapter 1.4 --- Classification of Diabetes Mellitus --- p.5 / Chapter 1.4.1 --- Type 1 Diabetes Mellitus --- p.5 / Chapter 1.4.2 --- Type 2 Diabetes Mellitus --- p.6 / Chapter 1.4.3 --- Other Specific Types --- p.7 / Chapter 1.4.4 --- Gestational Diabetes --- p.9 / Chapter 1.4.5 --- Clinical Stages of Diabetes --- p.9 / Chapter 1.5 --- Diagnostic Criteria of Diabetes Mellitus --- p.10 / Chapter 1.6 --- Complications of Diabetes Mellitus --- p.12 / Chapter 1.7 --- Pharmacological Treatment of Diabetes --- p.13 / Chapter 1.7.1 --- Treatment of Type 1 Diabetes --- p.13 / Chapter 1.7.2 --- Treatment of Type 2 Diabetes --- p.14 / Chapter 1.7.2.1 --- Sulphonylureas --- p.17 / Chapter 1.7.2.2 --- Biguanides --- p.18 / Chapter 1.7.2.3 --- α-Glucosidase Inhibitors --- p.19 / Chapter 1.7.2.4 --- Thiazolidinediones --- p.20 / Chapter 1.8 --- Diabetes and Traditional Chinese Medicine --- p.21 / Chapter 1.9 --- Project Objective --- p.27 / Chapter Chapter2 --- Botanical and Phytochemical Studies --- p.28 / Chapter 2.1 --- Introduction --- p.28 / Chapter 2.2 --- Materials --- p.31 / Chapter 2.3 --- Authentication of Herbal Material --- p.41 / Chapter 2.3.1 --- Materials --- p.41 / Chapter 2.3.2 --- Phytochemical Studies --- p.43 / Chapter 2.3.2.1 --- Sample Preparation --- p.43 / Chapter 2.3.2.2 --- Thin Layer Chromatography --- p.46 / Chapter 2.3.3 --- Results --- p.51 / Chapter 2.4 --- Extraction of Herbal Material --- p.56 / Chapter 2.4.1 --- Materials and Methods --- p.56 / Chapter 2.4.2 --- Results --- p.46 / Chapter 2.5 --- Quantification of Sugar Content in Herbal Extracts --- p.58 / Chapter 2.5.1 --- Introduction --- p.58 / Chapter 2.5.2 --- Materials and Methods --- p.58 / Chapter 2.5.3 --- Results --- p.61 / Chapter 2.6 --- Discussion --- p.65 / Chapter Chapter3 --- In vitro Studies on Formula 2 and its Individual Herbs --- p.68 / Chapter 3.1 --- Introduction --- p.68 / Chapter 3.2 --- Intestinal Glucose Absorption Studies --- p.69 / Chapter 3.2.1 --- Introduction --- p.69 / Chapter 3.2.2 --- Materials and Methods --- p.70 / Chapter 3.2.2.1 --- Preparation of BBMV --- p.71 / Chapter 3.2.2.2 --- BBMV Glucose Uptake Assay --- p.72 / Chapter 3.2.2.3 --- Bicinchoninic Acid (BCA) Protein Assay --- p.73 / Chapter 3.2.2.4 --- Preparation of Herbal Chloroform Extract --- p.74 / Chapter 3.2.2.5 --- Glucose Uptake Assay with Herbal Extracts --- p.75 / Chapter 3.2.3 --- Results --- p.76 / Chapter 3.3 --- Hepatic Gluconeogenesis Studies --- p.79 / Chapter 3.3.1 --- Introduction --- p.79 / Chapter 3.3.2 --- Materials and Methods --- p.82 / Chapter 3.3.2.1 --- Cell Culture --- p.83 / Chapter 3.3.2.2 --- Glucose Production Assay --- p.83 / Chapter 3.3.2.3 --- PEPCK Assay --- p.85 / Chapter 3.3.3 --- Results --- p.86 / Chapter 3.4 --- Cellular Glucose Uptake Studies --- p.88 / Chapter 3.4.1 --- Introduction --- p.88 / Chapter 3.4.2 --- Materials and Methods --- p.89 / Chapter 3.4.2.1 --- Cell Culture --- p.89 / Chapter 3.4.2.2 --- Differentiation of 3T3-L1 --- p.90 / Chapter 3.4.2.3 --- 2-Deoxy-D-glucose Uptake Assay --- p.91 / Chapter 3.4.3 --- Results --- p.92 / Chapter 3.5 --- Discussion --- p.96 / Chapter 3.5.1 --- Intestinal Glucose Absorption Studies by BBMV --- p.96 / Chapter 3.5.2 --- Hepatic Gluconeogenesis Studies by H4IIE Cells --- p.97 / Chapter 3.5.3 --- Cellular Glucose Uptake Studies by Hs68 and 3T3-L1 Cells --- p.99 / Chapter 3.5.4 --- Conclusions --- p.100 / Chapter Chapter4 --- In vivo Studies on Selected Herbs --- p.103 / Chapter 4.1 --- Introduction --- p.103 / Chapter 4.1.1 --- Animal Models of Type 2 Diabetes --- p.103 / Chapter 4.1.2 --- Chemically-induced Diabetic Models --- p.104 / Chapter 4.1.3 --- Neonatal-STZ Diabetic Rats --- p.107 / Chapter 4.2 --- Basal Glycaemia Test --- p.109 / Chapter 4.2.1 --- Animals --- p.109 / Chapter 4.2.2 --- Testing Method --- p.110 / Chapter 4.2.3 --- Results --- p.112 / Chapter 4.3 --- Oral Glucose Tolerance Test --- p.114 / Chapter 4.3.1 --- Animals --- p.114 / Chapter 4.3.2 --- Testing Method --- p.114 / Chapter 4.3.3 --- Results --- p.116 / Chapter 4.4 --- Discussion --- p.119 / Chapter Chapter5 --- Bioassay-guided Fractionation of Cortex Moutan --- p.125 / Chapter 5.1 --- Introduction --- p.125 / Chapter 5.1.1 --- Phytochemical Studies of Cortex Moutan --- p.125 / Chapter 5.2 --- Organic Extraction of Cortex Moutan --- p.128 / Chapter 5.2.1 --- Extraction Method --- p.128 / Chapter 5.2.2 --- Results --- p.129 / Chapter 5.3 --- BBMV Glucose Uptake Assay with Fraction CM C --- p.131 / Chapter 5.3.1 --- Materials and Methods --- p.131 / Chapter 5.3.2 --- Results --- p.131 / Chapter 5.4 --- In vivo Studies of Fraction CM-C --- p.133 / Chapter 5.4.1 --- Materials and Methods --- p.133 / Chapter 5.4.2 --- Results --- p.133 / Chapter 5.5 --- Fractionation of Fraction CM-C --- p.137 / Chapter 5.5.1 --- Materials and Methods --- p.137 / Chapter 5.5.2 --- Results --- p.139 / Chapter 5.6 --- BBMV Glucose Uptake Assay with CM-C Sub-fractions --- p.142 / Chapter 5.6.1 --- Results --- p.142 / Chapter 5.7 --- Isolation of Active Compound in Fraction CM-C4 --- p.144 / Chapter 5.7.1 --- Materials and Methods --- p.145 / Chapter 5.7.2 --- Results --- p.146 / Chapter 5.8 --- Structure Elucidation of CM-C4a --- p.148 / Chapter 5.8.1 --- Materials and Methods --- p.148 / Chapter 5.8.2 --- Results --- p.149 / Chapter 5.9 --- Effect of Paeonol in Oral Glucose Tolerance Test --- p.152 / Chapter 5.9.1 --- Materials and Methods --- p.152 / Chapter 5.9.2 --- Results --- p.153 / Chapter 5.10 --- Discussion --- p.155 / Chapter Chapter6 --- General Discussion --- p.163 / Chapter 6.1 --- Introduction --- p.163 / Chapter 6.2 --- Summary of Research Findings --- p.164 / Chapter 6.3 --- Limitations and Improvements --- p.167 / Chapter 6.4 --- Future Directions --- p.169 / Chapter 6.5 --- Conclusion --- p.170 / Appendices --- p.172 / Appendix 1 Low Resolution EI Mass Spectrum of Paeonol Reference --- p.173 / Appendix 2 Low Resolution EI Mass Spectrum of CM-C4a --- p.174 / Appendix 3 High Resolution EI Mass Spectrum of Paeonol Reference --- p.175 / Appendix 4 High Resolution EI Mass Spectrum of CM-C4a --- p.176 / Appendix 5 1H-NMR Spectrum of Paeonol Reference --- p.177 / Appendix 6 1H-NMR Spectrum of CM-C4a --- p.178 / References --- p.179

by Tsz-Wan Ho. / Thesis submitted in: July 2003. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2004. / Includes bibliographical references (leaves 107-122). / Abstracts in English and Chinese. / Acknowledgements --- p.i / Abstract --- p.ii / 摘要 --- p.iv / Content --- p.vi / Abbreviations --- p.x / List of Figures --- p.xi / List of tables --- p.xiv / Chapter CHAPTER 1 --- GENERAL INTRODUCTION --- p.1 / Chapter 1.1 --- Alzheimer'sDisease --- p.1 / Chapter 1.2 --- Hallmarks of AD --- p.3 / Chapter 1.2.1 --- The amyloid cascade hypothesis --- p.3 / Chapter 1.2.2 --- The tauopathy hypothesis --- p.4 / Chapter 1.3 --- The Cholinergic Hypothesis --- p.6 / Chapter 1.3.1 --- Cholinergic drug therapy --- p.7 / Chapter 1.3.2 --- Acetylcholinesterase inhibitors --- p.8 / Chapter 1.3.2.1 --- Tacrine --- p.10 / Chapter 1.3.2.2 --- Donepezil --- p.10 / Chapter 1.3.2.3 --- Rivastigimine - ENA-713 --- p.11 / Chapter 1.4 --- AChE inhibitors from plants --- p.12 / Chapter 1.4.1 --- Galanthamine --- p.12 / Chapter 1.4.2 --- Huperzine --- p.14 / Chapter 1.4.3 --- α-onocerin --- p.15 / Chapter 1.4.4 --- (+)-alpha-viniferin --- p.16 / Chapter 1.5 --- My project --- p.17 / Chapter CHAPTER 2 --- MATERIALS AND METHODS --- p.18 / Chapter 2.1 --- Preparation of CMM --- p.18 / Chapter 2.2.1 --- Selecting criteria and sources --- p.18 / Chapter 2.2.2 --- Preparation of aqueous extract --- p.18 / Chapter 2.2.3 --- Preparation of ethanol extract --- p.18 / Chapter 2.3 --- Routine maintenance of cell lines --- p.19 / Chapter 2.4 --- Toxicity test --- p.19 / Chapter 2.5 --- Ellman assay --- p.20 / Chapter 2.6 --- Ellman assay over BuChE --- p.21 / Chapter 2.7 --- Drugs --- p.21 / Chapter CHAPTER 3 --- SCREENING OF ACETYLCHOLINESTERASE INHIBITORS FROM CHINESE MEDICINAL MATERIALS --- p.23 / Chapter 3.1 --- Introduction --- p.23 / Chapter 3.2 --- Materials and Methods --- p.23 / Chapter 3.3 --- Results and discussion --- p.24 / Chapter 3.3.1 --- Preliminary screening of 45 selected TCMs for AChE inhibition --- p.24 / Chapter 3.3.2 --- Rescreening of drugs that show AChE inhibition in both aqueous and organic extracts --- p.25 / Chapter 3.4 --- Discussion --- p.28 / Chapter CHAPTER 4 --- CHARACTERIZATION OF ANTI-ACETYLCHOLINESTERASE ACTIVITY FROM SALVIA MBLTIORRHIZA BGE.(丹參) --- p.33 / Chapter 4.1 --- Introduction --- p.33 / Chapter 4.1.1 --- Clinical application of Danshen --- p.34 / Chapter 4.1.2 --- Pharmacological properties of Danshen and Salvia species --- p.34 / Chapter 4.1.2.1. --- Antiinflammatory and antibacterial responses --- p.35 / Chapter 4.1.2.2 --- Diabetes --- p.35 / Chapter 4.1.2.3 --- Alcoholism --- p.35 / Chapter 4.1.2.4 --- Apoptosis --- p.36 / Chapter 4.1.2.5 --- The effect of Salvia extracts on neuro-receptors --- p.36 / Chapter 4.1.3 --- Anti-cholinesterase activity by the Salvia species --- p.37 / Chapter 4.1.4 --- Active components from Salvia miltiorrhiza Bge --- p.38 / Chapter 4.2 --- Effects of tanshinone derivatives on AChE --- p.39 / Chapter 4.2.1 --- Materials and Methods --- p.39 / Chapter 4.2.2. --- Results --- p.39 / Chapter 4.3 --- Discussion --- p.50 / Chapter CHAPTER 5 --- EXTRACTION OF CRYPTOTANSHINONE FROM SALVIA MILTIORRHIZA --- p.54 / Chapter 5.1 --- Introduction --- p.54 / Chapter 5.1.1 --- Reverse phase high performance liquid chromatography (RP-HPLC) --- p.55 / Chapter 5.2 --- Materials and Methods --- p.56 / Chapter 5.2.1 --- Extracts of Danshen from different sources for obtaining the chemical profile --- p.55 / Chapter 5.2.2 --- Reverse phase high performance liquid chromatography (RP-HPLC) --- p.57 / Chapter 5.2.2.1 --- Analytical RP-HPLC --- p.57 / Chapter 5.2.2.2 --- Preparative RP-HPLC --- p.58 / Chapter 5.3 --- Results --- p.60 / Chapter 5.3.1 --- Identification of Peaks that contain the proposed active components --- p.60 / Chapter 5.3.2 --- Different samples of Danshen contain different amount of active components that can exert inhibitory effect on hAChE --- p.66 / Chapter 5.4 --- Discussion --- p.75 / Chapter CHAPTER 6 --- EFFECT OF CRYPTOTANSHINONE ON CALCIUM MOVEMENT in SH-SY5Y Cell --- p.80 / Chapter 6.1 --- Introduction --- p.80 / Chapter 6.2 --- Materials and Methods --- p.82 / Chapter 6.2.1 --- Reagents and drugs --- p.82 / Chapter 6.2.2 --- Calcium fluorimetry --- p.82 / Chapter 6.3 --- Results --- p.85 / Chapter 6.4 --- Discussion --- p.96 / Chapter CHAPTER 7 --- GENERAL DISCUSSION --- p.98 / Chapter 7.1 --- Structure-function relationship of crytotanshinone and dihydrotanshinone I --- p.98 / Chapter 7.2 --- Further study on cryptotanshinone and dihydrotanshinone I --- p.100 / Chapter 7.2.1 --- Modulation on nictonic receptor --- p.100 / Chapter 7.2.2 --- Behavioral study on mice --- p.101 / Chapter 7.2.3 --- Large scale production of the desired active components --- p.102 / Chapter 7.3 --- Study on other candidate herbs --- p.102 / References --- p.107

Chan Yiu-Pong. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2004. / Includes bibliographical references (leaves 98-112). / Abstracts in English and Chinese. / Abstract --- p.i / Acknowledgement --- p.v / Declaration --- p.vi / Table of content --- p.vii / List of Tables --- p.x / List of Figures --- p.xi / List of abbreviation --- p.xiv / Chapter Chapter 1 --- Introduction --- p.1 / Chapter 1.1 --- Introduction of Semen Oroxyli --- p.1 / Chapter 1.1.1 --- Chemical constituents of Oroxylum indicum seed --- p.3 / Chapter 1.1.2 --- Pharmacological studies --- p.3 / Chapter 1.2 --- Introduction to tracheal secretion --- p.6 / Chapter 1.2.1 --- Mucus composition --- p.6 / Chapter 1.2.2 --- Sputum formation --- p.6 / Chapter 1.2.3 --- Expectorant --- p.7 / Chapter 1.2.3.1 --- Secretolytic drugs --- p.7 / Chapter 1.2.3.2 --- Mucolytic drugs --- p.8 / Chapter 1.2.4 --- Assays of studying expectorant activity --- p.10 / Chapter 1.2.4.1 --- Tracheal phenol red secretion system --- p.10 / Chapter 1.3 --- Introduction to oxidant and antioxidant --- p.11 / Chapter 1.3.1 --- Oxidants/ reactive oxygen species --- p.11 / Chapter 1.3.1.1 --- Production of oxidants/ reactive oxygen species --- p.11 / Chapter 1.3.1.2 --- Reactive oxygen species reaction products --- p.12 / Chapter 1.3.1.2.1 --- DNA damage --- p.13 / Chapter 1.3.1.2.2 --- Protein damage --- p.13 / Chapter 1.3.1.2.3 --- Lipid damage --- p.14 / Chapter 1.3.2 --- Antioxidant --- p.14 / Chapter 1.3.2.1 --- Endogenous antioxidants --- p.15 / Chapter 1.3.2.1.1 --- Superoxide dismutase --- p.15 / Chapter 1.3.2.1.2 --- Catalase --- p.15 / Chapter 1.3.2.1.3 --- Glutathione and glutathione peroxidases --- p.15 / Chapter 1.3.3 --- Synthetic and natural antioxidants --- p.16 / Chapter 1.3.3.1 --- Vitamin C --- p.17 / Chapter 1.3.3.2 --- Tocopherols (Vitamin E) --- p.17 / Chapter 1.3.4 --- Assays for studying antioxidative activities --- p.19 / Chapter 1.3.4.1 --- DPPH radical scavenging system --- p.19 / Chapter 1.3.4.2 --- PMS-NADH system --- p.19 / Chapter 1.3.4.3 --- APPH-induced hemolysis system --- p.20 / Chapter 1.4 --- Objectives of the research --- p.22 / Chapter Chapter 2 --- Materials and Methods --- p.24 / Chapter 2.1 --- Materials --- p.24 / Chapter 2.1.1 --- Semen Oroxyli --- p.24 / Chapter 2.1.2 --- Animals --- p.24 / Chapter 2.1.3 --- Chemicals --- p.25 / Chapter 2.2 --- Methods --- p.28 / Chapter 2.2.1 --- Assay for studying expectorant activity --- p.28 / Chapter 2.2.1.1 --- Phenol red standard curve --- p.28 / Chapter 2.2.1.2 --- Tracheal phenol red secretion system --- p.28 / Chapter 2.2.2 --- Assays for studying antioxidative activity --- p.30 / Chapter 2.2.2.1 --- DPPH radicals scavenging system --- p.30 / Chapter 2.2.2.2 --- PMS-NADH system --- p.31 / Chapter 2.2.2.3 --- AAPH-induced red blood cell hemolysis system --- p.32 / Chapter 2.2.3 --- "Extraction, fractionation and purification of Semen Oroxyli" --- p.33 / Chapter 2.2.3.1 --- 70% ethanol extraction of Semen Oroxyli --- p.33 / Chapter 2.2.3.2 --- Fractionation in polyamide column --- p.33 / Chapter 2.2.3.3 --- Fractionation in resin column --- p.34 / Chapter 2.2.3.4 --- Sub-fractions separation from 95% ethanol soluble fraction --- p.34 / Chapter 2.2.3.5 --- Pure compounds obtained from sub-fractions --- p.34 / Chapter 2.2.4 --- Nulcear magnetic resonance (NMR) for identification --- p.38 / Chapter Chapter 3 --- Results of Expectorant Activity --- p.39 / Chapter 3.1 --- Expectorant Activity --- p.39 / Chapter 3.1.1 --- Expectorant Activity on Semen Oroxyli ethanol extract --- p.39 / Chapter 3.1.2 --- Expectorant activities of fractionations of Semen Oroxyli ethanol extract --- p.39 / Chapter Chapter 4 --- Results of Antioxidative Activity --- p.44 / Chapter 4.1 --- Antioxidative activity --- p.44 / Chapter 4.1.1 --- Antioxidative activity of 70% ethanol extract --- p.44 / Chapter 4.1.2 --- Antioxidative activity of fractions of 70% ethanol extract --- p.49 / Chapter 4.1.3 --- Antioxidative activity on sub-fractions fractionated from 95% ethanol-soluble fraction --- p.50 / Chapter 4.1.4 --- Antioxidative activity on pure compounds isolated from sub-fractions --- p.59 / Chapter Chapter 5 --- Results of Identification of Pure compounds --- p.68 / Chapter 5.1 --- Identification of compounds --- p.68 / Chapter 5.1.1 --- Compound A --- p.68 / Chapter 5.1.2 --- Compound B --- p.69 / Chapter 5.1.3 --- Compound C --- p.69 / Chapter 5.1.4 --- Compound D --- p.70 / Chapter 5.1.5 --- Compound E --- p.71 / Chapter Chapter 6 --- Discussion --- p.73 / Chapter 6.1 --- Discussion of expectorant activity of Semen Oroxyli --- p.73 / Chapter 6.2 --- Discussion of antioxidative activity of Semen Oroxyli --- p.75 / Chapter 6.3 --- General Discussion --- p.77 / Chapter Chapter 7 --- Conclusions --- p.82 / Appendix A Procedures for preparing the phenol red standard curve for tracheal phenol red secretion system. --- p.85 / Appedix B 1H NMR and 13C NMR spectra --- p.87 / References --- p.98

Ng, Chor Fung. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2007. / Includes bibliographical references (leaves 130-144). / Abstracts in English and Chinese. / ABSTRACT (ENGLISH VERSION) --- p.i / ABSTRACT (CHINESE VERSION) --- p.iv / ACKNOWLEDGEMENTS --- p.vi / TABLE OF CONTENTS --- p.viii / LIST OF TABLES AND FIGURES --- p.xii / ABBREVIATIONS --- p.xiv / Chapter CHAPTER 1. --- GENERAL INTRODUCTION --- p.1 / Chapter 1.1 --- Definition of asthma --- p.1 / Chapter 1.2 --- Asthma epidemiology --- p.2 / Chapter 1.3 --- Pathogenesis of Asthma --- p.3 / Chapter 1.3.1 --- Gene-environment interaction --- p.3 / Chapter 1.3.2 --- Allergens and atopic sensitization --- p.4 / Chapter 1.3.3 --- Other environmental factors --- p.5 / Chapter 1.4 --- House dust mite (HDM) --- p.5 / Chapter 1.4.1 --- Characteristics of HDM allergens --- p.5 / Chapter 1.4.2 --- HDM and asthma --- p.6 / Chapter 1.5 --- Pathophysiology of asthma --- p.8 / Chapter 1.5.1 --- Airway inflammation --- p.8 / Chapter 1.5.1.1 --- Cellular mechanism --- p.8 / Chapter 1.5.1.2 --- Characteristics of chronic inflammation --- p.9 / Chapter 1.5.1.3 --- Inflammatory cells in airway inflammation --- p.10 / Chapter 1.5.1.3.1 --- Mast cell --- p.10 / Chapter 1.5.1.3.2 --- Macrophages --- p.11 / Chapter 1.5.1.3.3 --- T lymphocytes --- p.12 / Chapter 1.5.1.3.4 --- Eosinophils --- p.12 / Chapter 1.5.1.3.5 --- Epithelial cells --- p.13 / Chapter 1.5.1.4 --- Cytokines in asthma --- p.14 / Chapter 1.5.1.4.1 --- Inflammatory cytokines --- p.14 / Chapter 1.5.1.4.1.1 --- Interleukin-4 --- p.14 / Chapter 1.5.1.4.1.2 --- Interleukin-5 --- p.14 / Chapter 1 5.1.4.1.3 --- Interleukin-6 --- p.15 / Chapter 1.5.1.4.1.4 --- Granulocyte Monocyte Colony Stimulating Factor (GM-CSF) --- p.15 / Chapter 1.5.1.4.1.5 --- Tumor Necrosis Factor-α (TNF-α) --- p.16 / Chapter 1.5.1.4.2 --- Anti-inflammatory cytokines --- p.17 / Chapter 1.5.1.4.2.1 --- Interleukin-10 --- p.17 / Chapter 1.5.1.4.2.2 --- Interferon-γ(IFN-γ) --- p.17 / Chapter 1.5.2 --- Airway hyperresponsiveness (AHR) --- p.18 / Chapter 1.5.3 --- A irway remodeling --- p.19 / Chapter 1.6 --- Asthma therapy --- p.21 / Chapter 1.6.1 --- β2-agonists --- p.21 / Chapter 1.6.2 --- Cromolyn and nedocromil --- p.21 / Chapter 1.6.3 --- Theophylline --- p.22 / Chapter 1.6.4 --- Leukotriene modifiers --- p.22 / Chapter 1.6.5 --- Corticosteroids --- p.23 / Chapter 1.7 --- Traditional Chinese Medicine --- p.24 / Chapter 1.7.1 --- Introduction --- p.24 / Chapter 1.7.2 --- Traditional Chinese Medicine (TCM) --- p.24 / Chapter 1.7.3 --- "Chinese herbal formula, CU Formula 2 (CUF2) and Kuan Dong Hua" --- p.26 / Chapter 1.8 --- Objectives of our studies --- p.28 / Chapter CHAPTER 2. --- ESTABLISHMENT OF A HDM-INDUCED ASTHMATIC ANIMAL MODEL IN SD RATS --- p.32 / Chapter 2.1 --- Introduction --- p.32 / Chapter 2.2 --- Materials and methods --- p.33 / Chapter 2.2.1 --- Buffers and solutions --- p.33 / Chapter 2.2.2 --- Animals --- p.33 / Chapter 2.2.3 --- Preparation of aluminum hydroxide gel --- p.34 / Chapter 2.2.4 --- HDMAllergen --- p.34 / Chapter 2.2.5 --- Sensitization Procedure --- p.35 / Chapter 2.2.6 --- Intratracheal instillation challenge --- p.35 / Chapter 2.2.7 --- Bronchoalveolar lavage (BAL) and BAL Cell counting --- p.36 / Chapter 2.2.8 --- Lung Histopathological Analysis --- p.37 / Chapter 2.2.9 --- Measurement of cytokine and chemokine by Enzyme-Linked Immunosorbent Assay (ELISA) --- p.39 / Chapter 2.2.10 --- Statistical Analysis --- p.40 / Chapter 2.3 --- Results --- p.41 / Chapter 2.3.1 --- Cellular Analysis of BALF --- p.41 / Chapter 2.3.2 --- Histopathology --- p.42 / Chapter 2.3.3 --- Cytokine and chemokine --- p.43 / Chapter 2.4 --- Discussion --- p.44 / Chapter CHAPTER 3. --- IMMUNOMODULATORY EFFECT OF CUF2 AND KUAN DONG HUA IN A RAT MODEL OF HDM-INDUCED ASTHMA --- p.65 / Chapter 3.1 --- Introduction --- p.65 / Chapter 3.2 --- Materials and methods --- p.67 / Chapter 3.2.1 --- Herbal materials and extraction method --- p.67 / Chapter 3.2.2 --- "Antigen sensitization, challenge, and treatment" --- p.68 / Chapter 3.2.3 --- Bronchoalveolar lavage and cell differential counts --- p.69 / Chapter 3.2.4 --- Histological Studies --- p.69 / Chapter 3.2.5 --- Measurement of BALF cytokines and chemokines --- p.70 / Chapter 3.2.6 --- "Body weight, thymus index and spleen index" --- p.70 / Chapter 3.2.7 --- Statistical analysis --- p.70 / Chapter 3.3 --- Results --- p.71 / Chapter 3.3.1 --- Effect of herbs and DXA on total cells and eosinophils in BALF --- p.71 / Chapter 3.3.2 --- Effect of herb and DXA on lung histology --- p.72 / Chapter 3.3.3 --- Effect of herbs and DXA on cytokine and chemokine level in BALF --- p.73 / Chapter 3.3.4 --- "Effect of herb and DXA on body weight, thymus index and spleen index" --- p.75 / Chapter 3.4 --- Discussion --- p.77 / Chapter CHAPTER 4. --- IMMUNOMODULATORY EFFECT OF KUAN DONG HUA ON HUMAN MAST CELLS (HMC-1) --- p.109 / Chapter 4.1 --- Introduction --- p.109 / Chapter 4.2 --- Materials and methods --- p.110 / Chapter 4.2.1 --- Reagents --- p.110 / Chapter 4.2.2 --- Cell line and Cell Culture --- p.111 / Chapter 4.2.3 --- Herb and extraction procedure --- p.111 / Chapter 4.2.4 --- Cell Viability Assay --- p.112 / Chapter 4.2.5 --- Assay of cytokine secretion --- p.113 / Chapter 4.2.6 --- Quantitative Analysis of cytokines --- p.113 / Chapter 4.2.7 --- Bacterial endotoxin contamination --- p.114 / Chapter 4.2.8 --- Statistical analysis --- p.115 / Chapter 4.3 --- Results --- p.116 / Chapter 4.3.1 --- Effect of Kuan Dong Hua on cell viability of HMC-I --- p.116 / Chapter 4.3.2 --- Effect of Kuan Dong Hua on cytokine release from HMC-I --- p.116 / Chapter 4.3.3 --- Effect of endotoxin contamination in the extract --- p.117 / Chapter 4.4 --- Discussion --- p.118 / Chapter CHAPTER 5. --- GENERAL CONCLUSION --- p.125 / Chapter 5.1 --- Conclusion --- p.125 / Chapter 5.2 --- Limitations of this study and Future work --- p.128 / REFERENCES --- p.130 / APPENDICES --- p.145 / Appendix A. Wright-Giemsa Stain for cytospin preparations --- p.145 / Appendix B. Hematoxylin & eosin (H&E) staining --- p.145 / Appendix C. Congo Red staining --- p.146 / Appendix D. Periodic acid-Schiff (PAS) staining --- p.146

published_or_final_version / Anatomy / Doctoral / Doctor of Philosophy

Thesis (MTech. degree in Pharmaceutical Sciences)--Tshwane University of Technology, 2008. / The aim of the study is to investigate whether the chemical composition and antimicrobial activity of orally administered medicinal plants are affected by in vitro dissolution and gastrointestinal absorption processes. Few in vitro screening assays for biological activities of plant extracts consider the effect of the gastrointestinal system. This is an important factor for the bioavailability of plant extracts intended to be administered via the oral route. In this study, crude water and methanol extracts of selected plants (green tea, 'buchu', thyme, sage and wild camphor) were prepared and exposed to simulated gastric fluid and simulated intestinal fluid during dissolution studies. The crude extracts and resulting simulated gastric fluid and simulated intestinal fluid products were screened for antimicrobial activity.

Diabetes mellitus (DM) has become a global threat in developing and developed countries, where diabetic patients are more prone to cardiovascular complications, a condition called diabetic cardiomyopathy. Studies have shown a direct link between hyperglycaemia and an increase in the production of reactive oxygen species in cardiac cells leading to diabetic cardiomyopathy. This study tests oleanolic acid, a bioactive compound from the plant Syzigium aromaticum as an antioxidant which could have a potential role in management of DM. Aims i) To extract Oleanolic acid (OA) from Syzigium aromaticim, ii) Investigate the antioxidant effects of plant derived OA in an in-vitro model of hyperglycaemia induced oxidative stress. Methods The flower buds of the Syzigium aromaticim [(Linnaeus) Merrill & Perry] (Myrtaceae) plant (commonly called cloves) were used to isolate OA. The ethyl acetate solubles from the cloves were subjected to chromatographic fractionation to yield OA powder. Spectroscopic analysis was done using 1D and 2D 1H and 13C NMR techniques for the identification of the structure of the compound. This compound was then used in vitro to test for its antioxidative properties. H9C2 cardiac myoblasts were employed which were treated with normoglycaemic (5.5 mM) and hyperglycaemic (33 mM) glucose conditions. The cells were then treated with oleanolic acid to test for its antioxidant properties. We looked at a dose-dependent (0, 20, 50 μM) and time-dependent effects of OA treatment (6 and 24 hrs) following 48 hours glucose exposure. ROS levels were measured using H2DCF-DA fluorescence staining using microscopy and flow cytometry techniques for analysis. xviii Results Recrystallisation of the powder with ethanol and inspection of the 1 and 2- dimensional 1H- and 13C-NMR spectra of the compound with comparison to literature data confirmed OA molecular structure and IUPAC numbering similar to that of literature characterized and confirmed the structure of oleanolic acid. In cell specific data high glucose treatments on H9C2 cells showed increased ROS production (22 ± 6 % and 20 ± 7 % n= 3 p< 0.01) for 6 and 24 hrs treatments, respectively, compared to their normoglycaemic control groups. The 6 h OA treated group showed a decrease in ROS production with 26.6 ± 17.4 % for the 20 μM while for 50 μM there was a 37.7 ± 14.3% decrease. A ROS reduction trend was observed in the normoglycaemic group, but this was significant at 24 hrs with 46.8 ± 45.3% and 57.3 ± 9 % for both 20 and 50 μM treatments, respectively. The 24 hrs OA treated group showed a dose-dependent decrease in ROS with 50 μM more pronounced (80.7% ± 4.5 %). The 20 μM OA treatments also showed a 15.7 ± 19 % decrease in ROS. Discussion In the present study, we have evaluated the antioxidant effects of OA in vitro following extraction of the compound from Syzigium aromaticim. The oxidative stress induced by hyperglycaemia was attenuated by oleanolic acid and this also translated into decreased ROS suggesting its use as an antioxidant in alleviating cardiovascular complications associated with diabetes mellitus.

<p>Infusions, decoctions, alcoholic preparations and other dosage forms of Artemisia afra are frequently used in South African traditional medicine. Generally when these preparations are made without applying good manufacturing practices they do not meet microbial quality control standards, safety and toxicity criteria and encourage poor patients compliance. To overcome the aforementioned disadvantages of traditional dosage forms a sold dosage form, i.e. a table might be recommended. The first objective of this study was to formulate and manufacture a rapid release tablet dosage of Artemisia afra that would contain an amount of plant material equivalent to that found in its traditional liquid dosage forms and that would meet conventional pharmaceutical standards. The second objective was to conduct a pilot study to obtain a preliminary profile of the bioavailability of select flavonoids presents in both the tablet and traditional liquid preparation of Artemisia afra in humans.</p>

Made available in DSpace on 2015-03-03T11:52:50Z (GMT). No. of bitstreams: 0 Previous issue date: 2014-07-25Bitstream added on 2015-03-03T12:06:59Z : No. of bitstreams: 1 000810985_20160725.pdf: 68863 bytes, checksum: e31a70f7392e0a8fa06a27c80bc87a8c (MD5) Bitstreams deleted on 2016-07-25T13:17:40Z: 000810985_20160725.pdf,. Added 1 bitstream(s) on 2016-07-25T13:18:47Z : No. of bitstreams: 1 000810985.pdf: 519868 bytes, checksum: 4fb63307e7d938ba410aa29f6f3812fb (MD5) / Sabe-se que dentro dos diferentes ecossistemas e em culturas de origem antrópica as plantas podem exercer interferência sobre outros vegetais ou microorganismos. Diferentes autores classificam esse evento como atividade alelopática, e esta ocorre principalmente pela liberação de biomoléculas (aleloquímicos), que podem variar sua constituição e classe química dependendo da espécie produtora. Estudos recentes têm demonstrado que os aleloquímicos constituem-se em importantes candidatos com potencial para serem utilizados como praguicidas e herbicidas naturais, transformando-se em aliados para o manejo agroecológico. Assim, o objetivo deste trabalho foi avaliar o potencial fitotóxico e antioxidante dos diferentes extratos das folhas de Tridax procumbens L., espécie ivasora e daninha, e Ouratea spectabilis (Mart. ex Engl.) Engl., nativa do cerrado brasileiro, por meio dos bioensaios laboratoriais de pré e pós- emergência em sementes de Lactuca sativa L. Além de avaliar o índice mitótico, aberrações cromossômicas e freqüência de micronúcleo em células de raiz de Allium cepa L. Adicionalmente, buscou-se determinar a atividade antioxidante e também elucidar fitoquimicamente os componentes fitotóxicos presentes nos extratos destas espécies. No bioensaio de pré-emergência as diferentes concentrações dos diferentes extratos reduziram os índices de germinação tanto para T. procumbens quanto para O. spectabilis quando comparados com o controle, alterando ainda o tempo médio, velocidade média e sincronismo da germinação. Semelhantemente ao encontrado nos bioensaios de pós- emergência, onde alterações no desenvolvimento de plântulas de alface (radícula e hipocótilo) foram observadas, apresentando inibição do crescimento da raíz principal e do hipocótilo quando comparados com o controle. Quanto ao índice mitótico, foi possível observar que os diferentes extratos... / It is known that within the different ecosystems and cultures of anthropogenic origin the plants can exercise interference on other plants or microorganisms. Different authors classify this event as allelopathic activity this occurs mainly by the release of biomolecules (allelochemicals) that can vary its constitution and chemical class depending on the producing species. Recent researches have shown that allelochemicals are important candidates with potential to be used as natural pesticides and herbicides, becoming an ally to agroecological management. Thus the aim of this study was to evaluate the phytotoxic and antioxidant potential of different extracts of leaves of Tridax procumbens L. ivasive and weed species and Ouratea spectabilis (Mart. ex Engl.) Engl. native to the Brazilian Cerrado through laboratory bioassays pre-and post-emergence in seeds of Lactuca sativa L. Besides assessing the mitotic index, chromosomal aberrations and frequency of micronuclei in root cells of Allium cepa L. We also determine the antioxidant activity and the chemical profile of phytotoxic compounds present in the extract of these species. In bioassay of pre-emergence the different concentrations of different extracts reduced the germination rates for both T. procumbens how O. spectabilis as compared with the control, even changing the average time, average speed and timing of germination. Similarly to that found in bioassays post-emergence, where changes in the development of lettuce seedlings (radicle and hypocotyl) were observed, with inhibition of root growth and hypocotyl compared with the control. Regarding the mitotic index was observed that the different extracts of the two species tested reduced when they were compared to the negative control. Introducing yet changes to the treatments with T. procumbens in rates of death and mutagenicity and O. spectabilis in the death rate, chromosomal alterations...

Orientador: Regildo Márcio Gonçalves da Silva / Banca: Anne Ligia Dokkedal Bosqueiro / Banca: Renata Giassi Udulutsch / Resumo: Sabe-se que dentro dos diferentes ecossistemas e em culturas de origem antrópica as plantas podem exercer interferência sobre outros vegetais ou microorganismos. Diferentes autores classificam esse evento como atividade alelopática, e esta ocorre principalmente pela liberação de biomoléculas (aleloquímicos), que podem variar sua constituição e classe química dependendo da espécie produtora. Estudos recentes têm demonstrado que os aleloquímicos constituem-se em importantes candidatos com potencial para serem utilizados como praguicidas e herbicidas naturais, transformando-se em aliados para o manejo agroecológico. Assim, o objetivo deste trabalho foi avaliar o potencial fitotóxico e antioxidante dos diferentes extratos das folhas de Tridax procumbens L., espécie ivasora e daninha, e Ouratea spectabilis (Mart. ex Engl.) Engl., nativa do cerrado brasileiro, por meio dos bioensaios laboratoriais de pré e pós- emergência em sementes de Lactuca sativa L. Além de avaliar o índice mitótico, aberrações cromossômicas e freqüência de micronúcleo em células de raiz de Allium cepa L. Adicionalmente, buscou-se determinar a atividade antioxidante e também elucidar fitoquimicamente os componentes fitotóxicos presentes nos extratos destas espécies. No bioensaio de pré-emergência as diferentes concentrações dos diferentes extratos reduziram os índices de germinação tanto para T. procumbens quanto para O. spectabilis quando comparados com o controle, alterando ainda o tempo médio, velocidade média e sincronismo da germinação. Semelhantemente ao encontrado nos bioensaios de pós- emergência, onde alterações no desenvolvimento de plântulas de alface (radícula e hipocótilo) foram observadas, apresentando inibição do crescimento da raíz principal e do hipocótilo quando comparados com o controle. Quanto ao índice mitótico, foi possível observar que os diferentes extratos... / Abstract: It is known that within the different ecosystems and cultures of anthropogenic origin the plants can exercise interference on other plants or microorganisms. Different authors classify this event as allelopathic activity this occurs mainly by the release of biomolecules (allelochemicals) that can vary its constitution and chemical class depending on the producing species. Recent researches have shown that allelochemicals are important candidates with potential to be used as natural pesticides and herbicides, becoming an ally to agroecological management. Thus the aim of this study was to evaluate the phytotoxic and antioxidant potential of different extracts of leaves of Tridax procumbens L. ivasive and weed species and Ouratea spectabilis (Mart. ex Engl.) Engl. native to the Brazilian Cerrado through laboratory bioassays pre-and post-emergence in seeds of Lactuca sativa L. Besides assessing the mitotic index, chromosomal aberrations and frequency of micronuclei in root cells of Allium cepa L. We also determine the antioxidant activity and the chemical profile of phytotoxic compounds present in the extract of these species. In bioassay of pre-emergence the different concentrations of different extracts reduced the germination rates for both T. procumbens how O. spectabilis as compared with the control, even changing the average time, average speed and timing of germination. Similarly to that found in bioassays post-emergence, where changes in the development of lettuce seedlings (radicle and hypocotyl) were observed, with inhibition of root growth and hypocotyl compared with the control. Regarding the mitotic index was observed that the different extracts of the two species tested reduced when they were compared to the negative control. Introducing yet changes to the treatments with T. procumbens in rates of death and mutagenicity and O. spectabilis in the death rate, chromosomal alterations... / Mestre